Ménard Delphine, Serk Henrik, Decou Raphaël, Pesquet Edouard
Umeå Plant Science Centre (UPSC), Department of Plant Physiology, Umeå University, 901 87, Umeå, Sweden.
Arrhenius Laboratories, Department of Ecology, Environment and Plant Sciences (DEEP), Svante Arrhenius väg 20A, Stockholm University, 160 91, Stockholm, Sweden.
Methods Mol Biol. 2017;1544:37-57. doi: 10.1007/978-1-4939-6722-3_4.
The development of inducible cell differentiation in suspension cultures led to multiple breakthroughs. It enabled the understanding of the chronology, duration, regulation and interdependency of the multiple events leading to fully functional specialized cells. The most studied cell differentiation in plants using inducible suspension cultures is the formation of tracheary elements (TEs) - the hydro-mineral sap conducting cells. Several in vitro systems established in different plant species have been developed to trigger TE formation on-demand. Here, we describe the establishment, harvesting and analysis of Arabidopsis thaliana stable habituated cell lines inducible by hormones to differentiate into TEs on-demand. Moreover, we explain the means to monitor and modify the chronology, duration and regulation of the progression of TE formation.
悬浮培养中诱导细胞分化的发展带来了多项突破。它使人们能够理解导致形成功能完备的特化细胞的多个事件的时间顺序、持续时间、调控及相互依存关系。利用诱导悬浮培养对植物中研究最多的细胞分化是管状分子(TEs)的形成,即传导水分和矿物质汁液的细胞。已开发出在不同植物物种中建立的多种体外系统,以按需触发TEs的形成。在此,我们描述了拟南芥稳定驯化细胞系的建立、收获及分析,这些细胞系可被激素诱导按需分化为TEs。此外,我们解释了监测和改变TEs形成进程的时间顺序、持续时间及调控的方法。